Affinage

RPE65

Retinoid isomerohydrolase · UniProt Q16518

Length
533 aa
Mass
60.9 kDa
Annotated
2026-04-28
100 papers in source corpus 25 papers cited in narrative 25 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

RPE65 is the retinoid isomerohydrolase of the vertebrate visual cycle, catalyzing the coupled hydrolysis and geometric isomerization of all-trans-retinyl esters to 11-cis-retinol — the rate-limiting step in regeneration of visual pigment chromophore for both rod and cone photoreceptors (PMID:16150724, PMID:16026160). Its catalytic mechanism depends on an iron-coordinating active site homologous to carotenoid cleavage dioxygenases, requires LRAT-generated retinyl ester substrate, and operates within a multiprotein complex on the ER membrane of RPE cells that includes RDH5 and RGR; membrane association is mediated by hydrophobic surface interactions and regulated S-palmitoylation at C112 and C146 (PMID:19805034, PMID:20100834, PMID:30914787). RPE65 is also expressed in green/red cone photoreceptors where it contributes to cone-autonomous chromophore supply (PMID:22171060, PMID:21753017). Loss-of-function mutations in RPE65 cause Leber congenital amaurosis, which is correctable by AAV-mediated gene replacement therapy that restores both rod and cone vision (PMID:18809924, PMID:16828753).

Mechanistic history

Synthesis pass · year-by-year structured walk · 13 steps
  1. 1997 Medium

    Establishing RPE65 as a participant in the retinoid visual cycle: its association with serum retinol-binding protein and 11-cis retinol dehydrogenase placed it at the interface of retinoid uptake and chromophore synthesis, though its enzymatic role was unknown.

    Evidence Co-immunoprecipitation from RPE extracts; liposome co-sedimentation and RET assays demonstrating hydrophobic membrane association without transmembrane domains

    PMID:9326941 PMID:9328280

    Open questions at the time
    • No direct enzymatic activity demonstrated for RPE65 itself
    • Co-IP associations not validated by reciprocal pulldown
    • Mechanism of membrane binding unresolved
  2. 2003 High

    RPE65 was shown to specifically bind and extract all-trans-retinyl esters from membranes and to reconstitute isomerase activity in rpe65−/− microsomes, establishing it as essential for — but not yet proven to be — the isomerase.

    Evidence Spectral-shift binding assay, gel filtration, FRET liposome assay, and isomerase reconstitution in rpe65−/− mouse microsomes

    PMID:14532273

    Open questions at the time
    • Could not distinguish whether RPE65 is the enzyme or a substrate-presenting chaperone
    • No active-site mutagenesis performed
  3. 2005 High

    Definitive identification of RPE65 as the isomerohydrolase itself: mutation of predicted iron-coordinating residues and iron chelation abolished catalytic activity, and LRAT co-expression was required for substrate provision, resolving the long-standing question of enzyme identity.

    Evidence Reconstitution in HEK293-F cells with HPLC retinoid analysis; active-site mutagenesis of iron-ligand residues; iron chelation; LCA mutations correlated with activity loss

    PMID:16150724

    Open questions at the time
    • No crystal structure yet available
    • Catalytic mechanism (concerted vs. stepwise hydrolysis/isomerization) unresolved
  4. 2005 High

    Quantitative in vivo analysis established RPE65 as the rate-limiting component for chromophore regeneration, with rhodopsin recovery following Michaelis kinetics as a function of RPE65 protein amount per eye.

    Evidence Quantitative RPE65 measurement across five mouse genotypes coupled with rhodopsin regeneration kinetics after photobleach

    PMID:16026160

    Open questions at the time
    • Whether RPE65 is also rate-limiting in cones was untested
    • Regulatory mechanisms controlling RPE65 abundance unknown
  5. 2006 High

    The Leu450Met RPE65 variant was identified as a genetic modifier of light-induced retinal degeneration in autosomal dominant RP, linking RPE65-dependent rhodopsin regeneration rate to photoreceptor susceptibility to phototoxicity.

    Evidence Epistasis analysis in transgenic RP mouse models with phototransduction and c-Fos knockouts; ERG and rhodopsin measurements

    PMID:16519667

    Open questions at the time
    • Whether the modifier effect extends to human RP patients unknown
    • Molecular mechanism by which L450M alters RPE65 kinetics not defined
  6. 2008 High

    Human gene therapy with AAV2-RPE65 restored both rod and cone vision in LCA patients, providing ultimate proof that RPE65 is required for chromophore production in both photoreceptor classes, while revealing abnormally slow rod dark adaptation post-treatment.

    Evidence Phase I/II clinical trial with psychophysical sensitivity measurements before and after subretinal AAV2-RPE65 injection

    PMID:18809924

    Open questions at the time
    • Cause of slow post-treatment rod resensitization kinetics unclear
    • Long-term durability of gene therapy unaddressed
  7. 2009 High

    The 2.14-Å crystal structure of bovine RPE65 revealed a large hydrophobic surface surrounding the active-site tunnel, explaining peripheral membrane association and providing a structural framework for mapping LCA mutations.

    Evidence X-ray crystallography of native bovine RPE65 with biophysical validation

    PMID:19805034

    Open questions at the time
    • No substrate-bound structure obtained
    • Catalytic mechanism of coupled hydrolysis-isomerization still unresolved at atomic level
  8. 2010 High

    RPE65 was found to operate within a multiprotein complex with RDH5 and RGR on RPE ER membranes, and its activity was shown to depend critically on intact phospholipid membrane structure.

    Evidence Co-immunoprecipitation of RPE65 with RDH5 and RGR from RPE microsomes; phospholipase A2 treatment abolishing isomerase activity; membrane extraction/phase separation

    PMID:20100834

    Open questions at the time
    • Stoichiometry and architecture of the complex undefined
    • Whether RGR directly modulates RPE65 catalysis unknown
  9. 2011 Medium

    RPE65 expression in cone photoreceptors (green/red but not blue cones in humans) was demonstrated to support cone-autonomous chromophore regeneration, expanding RPE65 function beyond the RPE.

    Evidence Immunohistochemistry on human and mouse retinas, in vitro ester hydrolysis in 661W cone cell line, ERG with exogenous chromophore across mouse strains

    PMID:21753017 PMID:22171060

    Open questions at the time
    • Relative quantitative contribution of cone RPE65 vs. RPE-derived chromophore not established
    • Mechanism regulating inverse RPE65 levels between RPE and cones unclear
  10. 2013 High

    FATP4 was identified as a negative regulator of RPE65 isomerase activity through substrate competition, with FATP4-deficient RPE showing enhanced chromophore regeneration and faster dark adaptation.

    Evidence cDNA library screen, isomerase assays in FATP4-KO vs. WT RPE, substrate competition with acyl-CoA species, ERG and rhodopsin regeneration in knockout mice

    PMID:23407971

    Open questions at the time
    • Physiological context for FATP4-mediated regulation (e.g., light/dark modulation) not defined
    • Whether ELOVL1-derived C24:0-CoA operates in vivo as an RPE65 inhibitor untested
  11. 2014 High

    PSMD13 was identified as a quality-control negative regulator that targets misfolded LCA mutant RPE65 for ubiquitin-proteasome degradation; chemical chaperones could rescue non-active-site but not active-site mutants, establishing a therapeutic strategy framework.

    Evidence Co-IP of PSMD13 with mutant RPE65, ubiquitination assay, proteasome inhibitor treatment, low-temperature and chemical chaperone rescue of isomerase activity

    PMID:24849605

    Open questions at the time
    • Whether PSMD13 interaction reflects direct binding or indirect proteasome engagement unclear
    • Clinical efficacy of chemical chaperone rescue not tested
  12. 2019 High

    S-palmitoylation at C112 and C146 was identified as a regulated post-translational modification essential for RPE65 ER membrane association and isomerase function, with LRAT dynamically modulating palmitoylation levels.

    Evidence Acyl-RAC palmitoylation assay, cysteine mutagenesis, pharmacological inhibition with 2-bromopalmitate/2-fluoropalmitate, membrane fractionation, isomerase activity assay

    PMID:30914787

    Open questions at the time
    • Identity of the palmitoyl acyltransferase(s) targeting RPE65 unknown
    • Structural basis for how palmitoylation complements hydrophobic surface-mediated membrane anchoring not resolved
  13. 2019 High

    The dominant D477G RPE65 mutation was shown to act primarily through aberrant mRNA splicing rather than through a missense protein effect, reframing pathogenic mechanism for this allele.

    Evidence CRISPR/Cas9 knock-in mouse model with RT-PCR splicing analysis; in vitro Exontrap assay validated for human c.1430A>G mutant

    PMID:30628748

    Open questions at the time
    • Whether splicing-corrective strategies (e.g., antisense oligonucleotides) can rescue function in vivo untested
    • Contribution of residual missense protein to dominant-negative toxicity not fully excluded

Open questions

Synthesis pass · forward-looking unresolved questions
  • Key unresolved questions include the atomic-level catalytic mechanism of coupled ester hydrolysis and retinoid isomerization, the identity of the palmitoyl acyltransferase(s) responsible for RPE65 palmitoylation, the stoichiometry and regulation of the RPE65-RDH5-RGR complex, and the quantitative contribution of cone-expressed RPE65 to diurnal vision.
  • No substrate-bound co-crystal structure available
  • Palmitoyl acyltransferase identity unknown
  • Complex stoichiometry and dynamics uncharacterized

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016787 hydrolase activity 8 GO:0016853 isomerase activity 4 GO:0008289 lipid binding 2
Localization
GO:0005783 endoplasmic reticulum 3 GO:0005886 plasma membrane 2
Pathway
R-HSA-1430728 Metabolism 6 R-HSA-9709957 Sensory Perception 4
Partners
FATP4LRATPSMD13RDH5RGR
Complex memberships
RPE65-RDH5-RGR visual cycle complex

Evidence

Reading pass · 25 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1997 RPE65 associates with serum retinol-binding protein and with the RPE-specific 11-cis retinol dehydrogenase, an enzyme active in the synthesis of the visual pigment chromophore 11-cis retinal, suggesting a role in the vitamin-A/retinoid cycle of the RPE. Co-immunoprecipitation / protein association assays Nature genetics Medium 9326941
2003 RPE65 specifically binds all-trans-retinyl palmitate (but not 11-cis-retinyl palmitate), extracts all-trans-retinyl esters from phospholipid membranes, and strongly stimulates enzymatic conversion of all-trans-retinyl palmitate to 11-cis-retinol in RPE microsomes; addition of RPE65 to rpe65-/- membranes (which have no detectable isomerase activity) restores isomerase activity to wild-type levels, suggesting RPE65 presents retinyl esters as substrate to the isomerase. Spectral-shift binding assay, gel filtration co-elution, co-immunoprecipitation, FRET liposome assay, in vitro isomerase activity assay with rpe65-/- mouse microsomes The Journal of biological chemistry High 14532273
2005 RPE65 is the isomerohydrolase of the visual cycle: mutations in predicted iron-coordinating residues (homologous to carotenoid oxygenase active-site residues) abolish 11-cis-retinoid production; iron chelation also abolishes activity; coexpression with LRAT is needed for high-level 11-cis-retinol production; disease-causing LCA mutations cause partial to total loss of isomerization activity proportional to clinical severity. Reconstitution of visual cycle in 293-F cells, HPLC retinoid analysis, active-site mutagenesis, iron chelation experiments Proceedings of the National Academy of Sciences of the United States of America High 16150724
2005 The rate of 11-cis-retinal production in vivo is a Michaelis function of RPE65 molecule quantity per eye (Vmax ~18 pmol/min, Km ~1.7 pmol), and at subsaturating RPE65 levels each molecule can deliver retinyl ester to the isomerohydrolase at ~10 molecules/min, establishing RPE65 as rate-limiting for chromophore synthesis. Quantitative biochemistry of RPE65 per eye across five mouse genotypes combined with in vivo rhodopsin regeneration kinetics after bleach Biochemistry High 16026160
2006 Point mutations Y144D and P363T (associated with Leber congenital amaurosis) significantly decrease RPE65 protein stability, alter its subcellular localization, and abolish its isomerohydrolase activity. Expression of mutant RPE65 in cells, Western blot stability assay, immunofluorescence localization, in vitro isomerase activity assay FEBS letters Medium 16828753
2008 RPE65 encodes the retinoid isomerase essential for 11-cis-retinal production; gene replacement restores both rod and cone photoreceptor-based vision in RPE65-LCA patients, but the reconstituted retinoid cycle shows abnormally slow rod resensitization kinetics (>8 h vs <1 h normal), while cone recovery is rapid. Clinical gene therapy trial with psychophysical visual sensitivity measurements before and after AAV2-RPE65 subretinal injection Proceedings of the National Academy of Sciences of the United States of America High 18809924
2009 Crystal structure of native bovine RPE65 at 2.14-Å resolution reveals a large lipophilic surface surrounding the entrance to the catalytic site that likely mediates membrane association with the ER; the structure provides a framework for understanding retinoid isomerization mechanism and how LCA-causing mutations disrupt protein function. X-ray crystallography of native bovine RPE65, supported by biophysical and biochemical validation Proceedings of the National Academy of Sciences of the United States of America High 19805034
2010 RPE65 enzymatic activity critically depends on membrane structural integrity: hydrophobic interactions dominate RPE65-membrane association; phospholipase A2 treatment of bovine RPE microsomes abolishes isomerase activity in parallel with phospholipid hydrolysis; RPE65 operates in a multiprotein complex with retinol dehydrogenase 5 (RDH5) and retinal G protein-coupled receptor (RGR) in RPE microsomes. Membrane extraction and phase separation experiments, phospholipase A2 treatment coupled to isomerase activity assays, co-immunoprecipitation of RPE65 with RDH5 and RGR from RPE microsomes The Journal of biological chemistry High 20100834
2002 RPE65 protein is expressed in mammalian cone photoreceptors (mouse, rabbit, cow) but not in rods, as demonstrated by immunohistochemistry on flatmounted retinas and double-labeling with PNA lectin; knockout of RPE65 abolishes this cone immunoreactivity, confirming the RPE65 gene identity. Immunohistochemistry on flatmounted and sectioned retinas, double-labeling with PNA lectin, RPE65 knockout controls Investigative ophthalmology & visual science High 11980880
2007 RPE65 is definitively localized by immunocytochemistry to the central RPE (not the neuroretina) in normal macaque, with immunoblotting showing ~4-fold higher retinoid isomerase activity in central vs. peripheral RPE; early cone photoreceptor loss in RPE65-LCA patients establishes that robust RPE65-based chromophore production is important for cone survival. Immunocytochemistry on primate RPE sections, immunoblotting, in vitro isomerase activity assay on central vs peripheral RPE fractions, in vivo microscopy of patient foveas Proceedings of the National Academy of Sciences of the United States of America High 17848510
2008 Novel Gly244Val RPE65 mutation close to the catalytic center causes protein instability in cultured cells and complete loss of RPE65-dependent isomerase activity, as shown by cell sorting and immunoblotting; homology modeling with the carotenoid oxygenase ACO template locates this and other inactivating mutations near the active site. Bicistronic expression with EGFP, flow cytometry/cell sorting, immunoblotting, enzymatic isomerase assay, structural homology modeling Biochemistry Medium 18722466
1997 RPE65 associates with phospholipid liposomes in a Ca2+-independent manner via hydrophobic interactions, and this association is responsible for its tight membrane co-sedimentation despite lacking transmembrane domains; the interaction causes phospholipid vesicle aggregation without fusion. Liposome co-sedimentation, resonance energy transfer (RET) assays, IAM.PC affinity binding Archives of biochemistry and biophysics Medium 9328280
1998 Translational regulation of RPE65 is mediated by a specific translation inhibitory element (TIE) located within 170 nucleotides downstream of the stop codon in the 3'-UTR, which requires interaction with the upstream RPE65 coding sequence or its product; the 5'-UTR does not affect translational efficiency. In vitro translation assays with RPE65 transcripts containing nested deletions of 5'- and 3'-UTRs; chloramphenicol acetyltransferase reporter heterologous control Archives of biochemistry and biophysics Medium 9721181
2001 RPE65 expressed in Sf9 insect cells exists in two forms: a non-membrane-associated form with mass close to calculated MW, and a membrane-associated form with significantly higher mass (~500 Da higher), indicating post-translational modification of the membrane-associated form. Baculovirus expression, affinity chromatography purification, MALDI mass spectrometry, Western blot, immunocytochemistry Investigative ophthalmology & visual science Medium 11381042
2006 The Rpe65 Leu450Met sequence variant acts as a genetic modifier of light-induced retinal degeneration in a transgenic mouse model of autosomal dominant RP: mice with Rpe65(450Met) show reduced degeneration and retain more rhodopsin, and the protective pathway involves phototransduction activity (not c-Fos), implicating regulation of daily photon absorption rate via rhodopsin regeneration kinetics as the mechanism. Genetic modifier analysis in transgenic mouse models, electroretinography, rhodopsin measurements, double-mutant epistasis with phototransduction and c-Fos knockouts The European journal of neuroscience High 16519667
2014 Three LCA-associated RPE65 missense mutations (L22P, T101I, L408P) at non-active sites cause misfolding and aggregation via disulfide bonds; the 26S proteasome subunit PSMD13 negatively regulates RPE65 by mediating rapid ubiquitination/proteasome-dependent degradation of these misfolded mutants; low-temperature rescue and chemical chaperones (sodium 4-phenylbutyrate, glycerol) restore activity of non-active-site mutants but not active-site mutants. Co-immunoprecipitation of PSMD13 with mutant RPE65, ubiquitination assay, proteasome inhibitor treatment, low-temperature rescue, isomerase activity assay, Western blot The Journal of biological chemistry High 24849605
2013 FATP4 (fatty acid transport protein 4) inhibits RPE65 isomerase activity by competing with RPE65 for its substrate (all-trans-retinyl esters); FATP4-deficient RPE shows significantly higher isomerase activity and faster 11-cis-retinaldehyde regeneration and rod light sensitivity recovery; ELOVL1-generated lignoceroyl (C24:0)-CoA inhibits 11-cis-retinol synthesis while palmitoyl (C16:0)-CoA promotes it. cDNA library screen, in vitro isomerase activity assay in FATP4-deficient vs wild-type RPE, substrate competition assay, electroretinography, rhodopsin regeneration kinetics in knockout mice The Journal of neuroscience : the official journal of the Society for Neuroscience High 23407971
2019 RPE65 is S-palmitoylated on residues C112 and C146 (approximately 25% of total RPE65); palmitoylation is required for ER membrane association and normal visual cycle function; inhibition of palmitoylation with 2-bromopalmitate or 2-fluoropalmitate completely abolishes membrane association; LRAT dynamically regulates RPE65 palmitoylation level (decreased in the presence of all-trans retinol). Acyl-RAC palmitoylation assay, mutational analysis of cysteine residues, pharmacological inhibition of palmitoylation, membrane association fractionation assay, isomerase activity assay Scientific reports High 30914787
2005 RPE65 surface epitopes co-elute with 11-cis retinol dehydrogenase (but not other visual cycle proteins) from immunoaffinity matrices prepared from solubilized bovine RPE membranes, suggesting a specific protein-protein interaction between RPE65 and 11-cis retinol dehydrogenase in the visual cycle complex. Immunoaffinity purification with monoclonal antibodies, Western blot identification of co-eluting proteins Molecular vision Low 16379027
2018 The dominant D477G RPE65 mutation generates an aggregation-prone surface that may induce cellular toxicity through abnormal complex formation; knock-in mice show ubiquitinated RPE65, reduced expression, retinyl ester accumulation, delayed rhodopsin regeneration, and diminished ERG responses; cell lines expressing D477G show comparable isomerase activity to wild-type, suggesting a toxic gain-of-function mechanism rather than simple loss of catalytic activity. D477G knock-in mouse model, immunoblot, retinoid HPLC analysis, ERG, in vitro isomerase assay in cell lines, structural analysis of RPE65 chimera, crystal packing analysis Human molecular genetics High 29659842
2016 The D477G dominant RPE65 mutation acts as a dominant-negative: heterozygous knock-in mice show slower kinetics of 11-cis-retinal regeneration after light exposure and lower A-wave recovery (delayed dark adaptation) compared to wild-type and RPE65 heterozygous knockout mice, despite similar stationary ERG amplitudes. D477G knock-in mouse model, HPLC retinoid analysis, ERG before and after photobleaching, comparison with heterozygous knockout The American journal of pathology High 28041994
2019 The c.1430A>G (D477G) RPE65 mutation primarily causes aberrant mRNA splicing by generating an ectopic splice site, dramatically disrupting RPE65 protein expression; this splicing defect was confirmed for the human RPE65 c.1430G mutant in an in vitro Exontrap assay, indicating the dominant pathogenesis is largely due to splicing disruption rather than a missense protein effect. CRISPR/Cas9 knock-in mouse model, RT-PCR splicing analysis, in vitro Exontrap assay for human mutant, visual function tests, light stress experiments Human mutation High 30628748
2011 RPE65 is expressed selectively in human green/red cones but absent from blue cones; in the 661W cone cell line, RPE65 mediates ester hydrolysis for photopigment synthesis in vitro, suggesting cone-expressed RPE65 supports diurnal vision. Immunohistochemistry on human retinal sections, Western blot, in vitro ester hydrolysis assay in 661W cone cell line The Journal of neuroscience : the official journal of the Society for Neuroscience Medium 22171060
2011 RPE65 is present in cones of the rod-dominant mouse retina and its level in cones is inversely related to RPE65 level in RPE; in BALB/c mice with undetectable cone RPE65, exogenous chromophore administration enhances cone light sensitivity ~3-fold (indicating chromophore deficiency), whereas C57BL/6 mice with cone RPE65 show no such enhancement, supporting a role for cone RPE65 in photopigment regeneration. Immunohistochemistry, Western blot of RPE vs cone fractions, ERG after exogenous chromophore administration, comparison across mouse strains The Journal of neuroscience : the official journal of the Society for Neuroscience Medium 21753017
2018 A novel non-retinoid RPE65 inhibitor CU239 selectively inhibits RPE65 isomerase activity (IC50 ~6 μM) by competing with the all-trans-retinyl ester substrate; systemic CU239 administration in mice delays chromophore regeneration after bleach and partially protects retina against high-intensity light damage. In vitro isomerase activity assay with IC50 determination, substrate competition assay, HPLC retinoid analysis in mice, ERG, histological assessment of light damage Biochimica et biophysica acta. Molecular basis of disease High 29684583

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2017 Efficacy and safety of voretigene neparvovec (AAV2-hRPE65v2) in patients with RPE65-mediated inherited retinal dystrophy: a randomised, controlled, open-label, phase 3 trial. Lancet (London, England) 1472 28712537
2008 Human gene therapy for RPE65 isomerase deficiency activates the retinoid cycle of vision but with slow rod kinetics. Proceedings of the National Academy of Sciences of the United States of America 579 18809924
1997 Mutations in RPE65 cause autosomal recessive childhood-onset severe retinal dystrophy. Nature genetics 543 9326941
1998 Mutations in the RPE65 gene in patients with autosomal recessive retinitis pigmentosa or leber congenital amaurosis. Proceedings of the National Academy of Sciences of the United States of America 385 9501220
2005 Mutation of key residues of RPE65 abolishes its enzymatic role as isomerohydrolase in the visual cycle. Proceedings of the National Academy of Sciences of the United States of America 363 16150724
2009 Human RPE65 gene therapy for Leber congenital amaurosis: persistence of early visual improvements and safety at 1 year. Human gene therapy 262 19583479
2000 Genetics and phenotypes of RPE65 mutations in inherited retinal degeneration. Investigative ophthalmology & visual science 226 11095629
2010 Leber congenital amaurosis due to RPE65 mutations and its treatment with gene therapy. Progress in retinal and eye research 204 20399883
2005 Retinal degeneration 12 (rd12): a new, spontaneously arising mouse model for human Leber congenital amaurosis (LCA). Molecular vision 189 15765048
2005 Gene therapy restores vision-dependent behavior as well as retinal structure and function in a mouse model of RPE65 Leber congenital amaurosis. Molecular therapy : the journal of the American Society of Gene Therapy 171 16223604
1998 Congenital stationary night blindness in the dog: common mutation in the RPE65 gene indicates founder effect. Molecular vision 154 9808841
2007 Human cone photoreceptor dependence on RPE65 isomerase. Proceedings of the National Academy of Sciences of the United States of America 138 17848510
2020 Clinical Perspective: Treating RPE65-Associated Retinal Dystrophy. Molecular therapy : the journal of the American Society of Gene Therapy 132 33278565
2011 A dominant mutation in RPE65 identified by whole-exome sequencing causes retinitis pigmentosa with choroidal involvement. European journal of human genetics : EJHG 131 21654732
2009 Crystal structure of native RPE65, the retinoid isomerase of the visual cycle. Proceedings of the National Academy of Sciences of the United States of America 128 19805034
2009 RPE65: role in the visual cycle, human retinal disease, and gene therapy. Ophthalmic genetics 123 19373675
2015 The Status of RPE65 Gene Therapy Trials: Safety and Efficacy. Cold Spring Harbor perspectives in medicine 122 25635059
2021 Perifoveal Chorioretinal Atrophy after Subretinal Voretigene Neparvovec-rzyl for RPE65-Mediated Leber Congenital Amaurosis. Ophthalmology. Retina 120 33838313
2017 Safety and Long-Term Efficacy of AAV4 Gene Therapy in Patients with RPE65 Leber Congenital Amaurosis. Molecular therapy : the journal of the American Society of Gene Therapy 108 29033008
2003 Rpe65 is a retinyl ester binding protein that presents insoluble substrate to the isomerase in retinal pigment epithelial cells. The Journal of biological chemistry 108 14532273
2006 Lentiviral gene transfer of RPE65 rescues survival and function of cones in a mouse model of Leber congenital amaurosis. PLoS medicine 102 17032058
2002 Identification of the RPE65 protein in mammalian cone photoreceptors. Investigative ophthalmology & visual science 98 11980880
2018 The Natural History of Inherited Retinal Dystrophy Due to Biallelic Mutations in the RPE65 Gene. American journal of ophthalmology 96 30268864
2003 In vivo gene therapy in young and adult RPE65-/- dogs produces long-term visual improvement. The Journal of heredity 92 12692160
2018 Gene therapy for RPE65-related retinal disease. Ophthalmic genetics 91 30335549
2019 CRISPR-Cas9-mediated therapeutic editing of Rpe65 ameliorates the disease phenotypes in a mouse model of Leber congenital amaurosis. Science advances 89 31692906
2008 A comprehensive clinical and biochemical functional study of a novel RPE65 hypomorphic mutation. Investigative ophthalmology & visual science 74 18599565
2021 Retinal pigment epithelium 65 kDa protein (RPE65): An update. Progress in retinal and eye research 73 34607013
2019 Long-Term Structural Outcomes of Late-Stage RPE65 Gene Therapy. Molecular therapy : the journal of the American Society of Gene Therapy 62 31604676
2015 Comprehensive genotyping reveals RPE65 as the most frequently mutated gene in Leber congenital amaurosis in Denmark. European journal of human genetics : EJHG 62 26626312
2010 Self-complementary AAV-mediated gene therapy restores cone function and prevents cone degeneration in two models of Rpe65 deficiency. Gene therapy 62 20237510
2007 RDH12 and RPE65, visual cycle genes causing leber congenital amaurosis, differ in disease expression. Investigative ophthalmology & visual science 61 17197551
2010 Importance of membrane structural integrity for RPE65 retinoid isomerization activity. The Journal of biological chemistry 59 20100834
2010 Gene therapy in the second eye of RPE65-deficient dogs improves retinal function. Gene therapy 59 20703309
2005 Mole quantity of RPE65 and its productivity in the generation of 11-cis-retinal from retinyl esters in the living mouse eye. Biochemistry 59 16026160
2001 Expression, purification, and MALDI analysis of RPE65. Investigative ophthalmology & visual science 58 11381042
2012 RPE65 gene therapy slows cone loss in Rpe65-deficient dogs. Gene therapy 54 22951453
2010 Membrane-binding and enzymatic properties of RPE65. Progress in retinal and eye research 53 20304090
2020 The effect of human gene therapy for RPE65-associated Leber's congenital amaurosis on visual function: a systematic review and meta-analysis. Orphanet journal of rare diseases 52 32059734
2006 Rpe65 as a modifier gene for inherited retinal degeneration. The European journal of neuroscience 51 16519667
2018 Results at 5 Years After Gene Therapy for RPE65-Deficient Retinal Dystrophy. Human gene therapy 47 29869534
2007 Evidence for RPE65-independent vision in the cone-dominated zebrafish retina. The European journal of neuroscience 46 17868371
2018 Cost Utility of Voretigene Neparvovec for Biallelic RPE65-Mediated Inherited Retinal Disease. Value in health : the journal of the International Society for Pharmacoeconomics and Outcomes Research 45 30711060
2002 Clinical course and visual function in a family with mutations in the RPE65 gene. Archives of ophthalmology (Chicago, Ill. : 1960) 44 11786058
2002 Retinal degeneration and RPE transplantation in Rpe65(-/-) mice. Investigative ophthalmology & visual science 44 12356839
2020 Voretigene neparvovec-rzyl for treatment of RPE65-mediated inherited retinal diseases: a model for ocular gene therapy development. Expert opinion on biological therapy 42 32149547
2006 Visual cycle protein RPE65 persists in new retinal cells during retinal regeneration of adult newt. The Journal of comparative neurology 42 16485283
2009 Evaluation of 9-cis-retinyl acetate therapy in Rpe65-/- mice. Investigative ophthalmology & visual science 41 19407008
2009 Predicting the pathogenicity of RPE65 mutations. Human mutation 41 19431183
1998 Promoter analysis of RPE65, the gene encoding a 61-kDa retinal pigment epithelium-specific protein. Investigative ophthalmology & visual science 41 9501877
2014 Rescue of enzymatic function for disease-associated RPE65 proteins containing various missense mutations in non-active sites. The Journal of biological chemistry 40 24849605
2002 Prenatal human ocular degeneration occurs in Leber's congenital amaurosis (LCA2). The journal of gene medicine 40 12124981
1998 Cloning and localization of RPE65 mRNA in salamander cone photoreceptor cells1. Biochimica et biophysica acta 40 9838153
2011 RPE65 is present in human green/red cones and promotes photopigment regeneration in an in vitro cone cell model. The Journal of neuroscience : the official journal of the Society for Neuroscience 39 22171060
1997 RPE65, the major retinal pigment epithelium microsomal membrane protein, associates with phospholipid liposomes. Archives of biochemistry and biophysics 38 9328280
2023 Single Center Experience with Voretigene Neparvovec Gene Augmentation Therapy in RPE65 Mutation-Associated Inherited Retinal Degeneration in a Clinical Setting. Ophthalmology 37 37704110
2013 Persistence of non-viral vector mediated RPE65 expression: case for viability as a gene transfer therapy for RPE-based diseases. Journal of controlled release : official journal of the Controlled Release Society 37 24035979
2022 Epidemiology of Mutations in the 65-kDa Retinal Pigment Epithelium (RPE65) Gene-Mediated Inherited Retinal Dystrophies: A Systematic Literature Review. Advances in therapy 36 35098484
2022 Inflammation after Voretigene Neparvovec Administration in Patients with RPE65-Related Retinal Dystrophy. Ophthalmology 36 35760216
2018 Insights into the pathogenesis of dominant retinitis pigmentosa associated with a D477G mutation in RPE65. Human molecular genetics 35 29659842
2014 Pseudo-fovea formation after gene therapy for RPE65-LCA. Investigative ophthalmology & visual science 34 25537204
2013 Mutational screening of LCA genes emphasizing RPE65 in South Indian cohort of patients. PloS one 34 24066033
2008 Impact of retinal disease-associated RPE65 mutations on retinoid isomerization. Biochemistry 34 18722466
2013 Fatty acid transport protein 4 (FATP4) prevents light-induced degeneration of cone and rod photoreceptors by inhibiting RPE65 isomerase. The Journal of neuroscience : the official journal of the Society for Neuroscience 33 23407971
2018 Two-year progression analysis of RPE65 autosomal dominant retinitis pigmentosa. Ophthalmic genetics 32 29947567
2013 Successful gene therapy in older Rpe65-deficient dogs following subretinal injection of an adeno-associated vector expressing RPE65. Human gene therapy 32 24028205
2021 RPE65-associated inherited retinal diseases: consensus recommendations for eligibility to gene therapy. Orphanet journal of rare diseases 31 34088339
2019 Generation and Characterization of Induced Pluripotent Stem Cells and Retinal Organoids From a Leber's Congenital Amaurosis Patient With Novel RPE65 Mutations. Frontiers in molecular neuroscience 31 31572124
2021 Inherited Retinal Diseases Due to RPE65 Variants: From Genetic Diagnostic Management to Therapy. International journal of molecular sciences 30 34281261
2007 Electroretinographic analyses of Rpe65-mutant rd12 mice: developing an in vivo bioassay for human gene therapy trials of Leber congenital amaurosis. Molecular vision 30 17960108
2006 Phenotype of three consanguineous Tunisian families with early-onset retinal degeneration caused by an R91W homozygous mutation in the RPE65 gene. Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie 28 16518657
2005 RPE65 surface epitopes, protein interactions, and expression in rod- and cone-dominant species. Molecular vision 28 16379027
1999 Identification of RPE65 in transformed kidney cells. FEBS letters 28 10386590
2016 A Dominant Mutation in Rpe65, D477G, Delays Dark Adaptation and Disturbs the Visual Cycle in the Mutant Knock-In Mice. The American journal of pathology 27 28041994
2006 Cortical visual function in the rd12 mouse model of Leber Congenital Amarousis (LCA) after gene replacement therapy to restore retinal function. Vision research 27 16814838
2023 Classification and Growth Rate of Chorioretinal Atrophy after Voretigene Neparvovec-Rzyl for RPE65-Mediated Retinal Degeneration. Ophthalmology. Retina 26 37660736
2006 Impacts of two point mutations of RPE65 from Leber's congenital amaurosis on the stability, subcellular localization and isomerohydrolase activity of RPE65. FEBS letters 26 16828753
2020 Voretigene Neparvovec: A Review in RPE65 Mutation-Associated Inherited Retinal Dystrophy. Molecular diagnosis & therapy 25 32535767
2023 Outcomes and Adverse Effects of Voretigene Neparvovec Treatment for Biallelic RPE65-Mediated Inherited Retinal Dystrophies in a Cohort of Patients from a Single Center. Biomolecules 24 37892166
2019 Aberrant RNA splicing is the major pathogenic effect in a knock-in mouse model of the dominantly inherited c.1430A>G human RPE65 mutation. Human mutation 24 30628748
2011 Regeneration of photopigment is enhanced in mouse cone photoreceptors expressing RPE65 protein. The Journal of neuroscience : the official journal of the Society for Neuroscience 23 21753017
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2000 A homozygous deletion in RPE65 in a small Sardinian family with autosomal recessive retinal dystrophy. Molecular vision 22 11062306
1998 Role of the 3'-untranslated region of RPE65 mRNA in the translational regulation of the RPE65 gene: identification of a specific translation inhibitory element. Archives of biochemistry and biophysics 22 9721181
2022 Human primitive mesenchymal stem cell-derived retinal progenitor cells improved neuroprotection, neurogenesis, and vision in rd12 mouse model of retinitis pigmentosa. Stem cell research & therapy 21 35395806
2020 Cost Effectiveness of Voretigene Neparvovec for RPE65-Mediated Inherited Retinal Degeneration in Germany. Translational vision science & technology 21 32879773
2019 RPE65 mutation frequency and phenotypic variation according to exome sequencing in a tertiary centre for genetic eye diseases in China. Acta ophthalmologica 21 31273949
2005 Function of the protein RPE65 in the visual cycle. Nutrition reviews 21 15825812
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2019 The dual roles of RPE65 S-palmitoylation in membrane association and visual cycle function. Scientific reports 20 30914787
2018 A Cross-Sectional and Longitudinal Study of Retinal Sensitivity in RPE65-Associated Leber Congenital Amaurosis. Investigative ophthalmology & visual science 20 30025081
2004 RPE65 of retinal pigment epithelium, a putative receptor molecule for plasma retinol-binding protein, is expressed in human keratinocytes. The Journal of investigative dermatology 20 15009723
2018 A novel RPE65 inhibitor CU239 suppresses visual cycle and prevents retinal degeneration. Biochimica et biophysica acta. Molecular basis of disease 19 29684583
2018 Examining the Role of Cone-expressed RPE65 in Mouse Cone Function. Scientific reports 19 30242264
2010 Cleavage of the retinal pigment epithelium-specific protein RPE65 under oxidative stress. International journal of biological macromolecules 19 20510285
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2012 Vitamin A metabolism in benign and malignant melanocytic skin cells: importance of lecithin/retinol acyltransferase and RPE65. Journal of cellular physiology 18 21465477